Tests to inject live bird flu in humansVirus is weakened and genetically altered; technique offers promise for future diseases

THE ASSOCIATED PRESSMonday, December 19, 2005

WASHINGTON

In an isolation ward of a Baltimore hospital, up to 30 volunteers will participate in a bold experiment: A vaccine made with a live version of the most notorious bird flu will be sprayed into their noses.

First, scientists are dripping that vaccine into the tiny nostrils of mice. It doesn't appear harmful - researchers have weakened and genetically altered the virus so that no one should get sick or spread germs - and it protects the animals enough to try in people.

This is essentially FluMist for bird flu, and the hope is that, in the event of a flu pandemic, immunizing people through their noses could provide faster, more effective protection than the troublesome shots - made with a killed virus - the nation now is struggling to produce.

And if it works, this new vaccine frontier may not just protect against the bird-flu strain, called H5N1, considered today's top health threat. It offers the potential for rapid, off-the-shelf protection against whatever novel variation of the constantly evolving influenza virus shows up next - through a library of live-virus nasal sprays that the National Institutes of Health plans to freeze.

"It's high-risk, high-reward" research, said Dr. Brian Murphy, who heads the NIH laboratory where Dr. Kanta Subbarao is brewing the nasal sprays - including one for a different bird-flu strain that appeared safe during the first crucial human testing last summer.

"It might fail, but if it's successful, it might prevent hundreds of thousands of cases" of the next killer flu, Murphy said.

FluMist is the nation's nasal-spray vaccine that prevents regular winter flu. Developed largely through Murphy's lab, it's the only flu vaccine made with live but weakened influenza viruses.

The new project, a collaboration with FluMist manufacturer MedImmune Inc., piggybacks cutting-edge genetics technology onto that vaccine to create a line of FluMist-like sprays against different bird flus.

"That is a great, great idea," said Dr. John Treanor of the University of Rochester, among the flu specialists closely watching the project.

Regular winter flu shots are made with killed influenza viruses, and the government is stockpiling experimental bird-flu vaccine made the same way. But those bird-flu shots don't work as well as hoped. They require an incredibly high dose, delivered in two separate injections, to cause a protective immune response in people.

"In theory, a live-virus vaccine might actually work better. We don't know that because we've never tried one before," Treanor said.

Influenza is like a magician, constantly changing its clothes to avoid detection, making it difficult to develop effective vaccines.

Studding the virus' surface are two proteins called hemagglutinin - the H in H5N1 - and neuraminidase, the N. They act as a wardrobe: There are 16 known hemagglutinin versions, and nine neuraminidases.

They're also what prompts the immune system to make an attack, particularly hemagglutinin, the protein the body aims for when it makes flu-fighting antibodies.

When people catch the flu, they usually get H1 or H3 flu strains, which their bodies can recognize because variations have circulated among humans for many years.

Occasionally, genetically unique strains emerge. Until 1997, H5 strains had never been seen outside of birds. The virus essentially put on a coat that human immune systems didn't recognize. The result: Since 2003, a particularly strong H5N1 strain has infected more than 130 people in Asia, killing at least 70.

H9 and H7 strains have also recently jumped from birds to people, although so far they haven't been nearly as dangerous.

Researchers hope to create at least one live-virus nasal spray for each "H" subtype, a project costing about $16 million of the NIH's annual $67 million budget for flu vaccine research.

"The hemagglutinin is the major protective antigen, so that is what we're focusing on," explained Subbarao, a molecular geneticist who heads the project.

First on her list are the riskiest known bird flus: H5N1, with human tests planned for April; H9N2, which recently underwent the first round of human testing in an isolation ward at Johns Hopkins Bayview Medical Center; then an H7 strain, followed by an H6 strain believed to share genes with the H5N1.

"By no means are we confident we're picking the right strain" to make first because flu mutates so easily, Subbarao said.

Subbarao must customize the strains for safe vaccination: First, using a new technique called reverse genetics, she selects genes for bird-flu H and N antigens and removes genetic segments that make them dangerous. Then she adds the remaining gene segments to the regular weakened FluMist virus.

Stocks of the custom virus are grown in fertilized chicken eggs. Each is then carefully cracked by hand to drain out virus-loaded liquid that in turn is purified and put into a nasal spray.

In a high-security section of the lab, Subbarao wears a biohazard suit and exposes vaccinated mice to various bird-flu strains.

Then it's time for human testing - in a hospital isolation ward just in case the weakened virus could infect someone.

It shouldn't because "those problems don't exist in FluMist," said Murphy, citing studies of regular FluMist in day-care centers where youngsters routinely pass viruses back and forth.

Some studies have found that people can shed virus shortly after receiving regular FluMist. But, "to spread infection, you'd need much more (virus) than replicates in the nose," he said.

Hopkins researchers gave the first of Subbarao's vaccine candidates - the H9N2 spray - to 30 volunteers last summer. To be sure they couldn't spread the virus by coughing or sneezing, the volunteers underwent daily tests of their noses and throats.